Figure 3.

Detection of HLA-G1 proteins in JEG-3 cells but not in IGR and M8 melanoma cells by W6/32 mAb reacting against β2-microglobulin-associated HLA-A,-B, -C, and -G HC. (A) Immunoprecipitation of metabolically labeled proteins. Cells were metabolically labeled with 0.5 mCi of [³⁵S] methionine plus [³⁵S] cysteine for 1 h, and cell lysates were precipitated with W6/32, analyzed by SDS 12%/PAGE and autoradiographed. (B) Biotinylated cell surface proteins of melanoma and the JEG-3 cell lines were immunoprecipitated with W6/32 mAb, and immunoprecipitates then were separated by SDS 12%/PAGE and transferred to nitrocellulose membranes. Surface class I molecules were detected by using peroxidase-conjugated streptavidin.